LOCUS       sequence-354351-        6833 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier A0A0U5NXR6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6833)
  AUTHORS   Rembeza E, Engqvist MKM
  TITLE     Experimental and computational investigation of enzyme functional 
            annotations uncovers misannotation in the EC 1.1.3.15 enzyme class.
  JOURNAL   PLoS Comput Biol. 2021 Sep 23;17(9):e1009446. doi: 
            10.1371/journal.pcbi.1009446. eCollection 2021 Sep.
  PUBMED    34555022
REFERENCE   2  (bases 1 to 6833)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS Comput
            Biol."; date: "2021-09-23"; volume: "17(9):e1009446. doi"; pages: " 
            10.1371/journal.pcbi.1009446. eCollection 2021 Sep"
COMMENT     Sequence Label: A0A0U5NXR6
FEATURES             Location/Qualifiers
     source          1..6833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1553..1582
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1635..1653)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1649..1696
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1733..2188
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1820..1839)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2030..2051
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2214..2318
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2319..2387,2388..3179)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2537..2556)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3350..3938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3839..3858
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4092..4109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4124..4263
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4189..4208)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4349..4371
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4365..4556)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5365..6447)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6409..6428
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6448..6525)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6731..6750
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gaactcgtac gatgtagttg ttattggcgc
      121 gggcgtagtc ggttgcgcga tcgcgcgtga attgtcacgt tacaaactgc gcatttgtgt
      181 actggagaag gaacctgatg tagcttacgg taatagctct cgtaacacgg gaatgctgca
      241 tgcaggtttt acctacaagc ctgggtctct gaaggcggtg tgcgcagtgg aagggaatcg
      301 cgagtttgat caggttgcgc gtgagctcga tgtgccgttt cgtcgcagcg gtaaagttgt
      361 agttggtttt accgacaccg attataccaa cattctgaag tacaagtcaa ttggagaggt
      421 aaacggggtt gatggcttgg agattatcga taagaagaag ttaaaccaga ttgaacctaa
      481 tgcgcgtggt gaatttgcca tgtactcccc gatgtcaggt atcctggacc cgattcagta
      541 caccattgct ctggccgaga atgcccaaca gaatggtgta cacttttgtt ttaaccgcga
      601 agttcttggg ctgaaaacgg atacgtccca gtaccacatc actactaata atggcatggt
      661 gacttcgcgt tgggtgatta attcagcagg gatgtatagt acaaagattt cggacatgct
      721 gggtatcacc ggatatgtta cgcgcggttt taaaggtgaa tactatgtgc tggacaagaa
      781 ggcaggtatc ggccttcgca cacctgtgta tccggccccg aacgaaaagg gtggcttcgc
      841 gacgcacgcc accatcacta tcgatggcaa cgtgctggtg ggccctgata gttatatgac
      901 tgaaggccgc gaagattatt ccacaacacg tgaacatctg gcgggtctgg ttaaggactc
      961 ttcccaaatg tttgacaagg ttcgtccgga attctttatc aagacattcg cgggcatccg
     1021 ctggaagttg gttgatcccg acaccggcga ggttcaggac ttcgtgttag agcgtcgtga
     1081 ggacgctccg catgcgatta acctcgttgg cattgaaagt ccgggtatta cttgtgcact
     1141 cccgctggca cgccgtgttg tgcaaaagat gcaagaggta gatagcctga tttcgaatcc
     1201 agacttctgt ccctaccgca aaggtattac gcgctttagc gagctgagcc tggaagagaa
     1261 agagaaatta gtgcaggaca accctctgta cggtgagatt gtttgtcgtt gtgaaagcgt
     1321 taccaaagcg gaaatcctgc aagcgatcca taaccccctg ggtgcgcaca ccgtggcgtc
     1381 aatcaagaat cgtacccgtg ctactatggg ccgctgtcag ggtggctact gcgaaacccg
     1441 catcactaaa atgatccagg aagagctgaa tatcccggaa aaggaaattc tttatcagca
     1501 taaggactct tatatgttcg ttggcaaggt gcgtggcgcg gccgcactcg agcatcatca
     1561 ccatcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga
     1621 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     1681 cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc
     1741 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1801 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     1861 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     1921 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     1981 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2041 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2101 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2161 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     2221 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2281 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2341 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2401 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     2461 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     2521 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     2581 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     2641 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     2701 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     2761 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     2821 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt
     2881 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     2941 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3001 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3061 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3121 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3181 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     3241 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     3301 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3361 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3421 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3481 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3541 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3601 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3661 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3721 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3781 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3841 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3901 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3961 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     4021 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     4081 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     4141 tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc
     4201 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     4261 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4321 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4381 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat
     4441 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat
     4501 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat
     4561 gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg
     4621 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt
     4681 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag
     4741 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag
     4801 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg
     4861 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
     4921 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
     4981 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc catgccggcg
     5041 ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg
     5101 agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga
     5161 aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg
     5221 ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag
     5281 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     5341 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5401 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     5461 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     5521 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     5581 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     5641 cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     5701 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     5761 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     5821 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     5881 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     5941 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     6001 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     6061 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     6121 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     6181 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     6241 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     6301 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     6361 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     6421 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     6481 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     6541 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     6601 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     6661 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     6721 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     6781 tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aat
//
